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. 2016 Mar 7;7(15):20597–20611. doi: 10.18632/oncotarget.7972

Figure 5. MEK1 maintains liver CSC self-renewal dependent on SIRT1.

Figure 5

Non-CSCs (Huh7-NanogNeg cells) were prepared with overexpression SIRT1, and grown with 5 μM U0126 or DMSO for 48 hours. Liver CSCs (Huh7-NanogPos cells) co-cultured with 5 μM U0126 for 24 hours previously and overexpression SIRT1 for next 24 hours. Colony analysis of CSCs (C) and non-CSCs (A) which were cultured for 14 days and stained with crystal violet. Sphere analysis of CSCs (D) and non-CSCs (B) which were cultured for 7 days in non-adhesive culture system. All counting were performed in triplicate. (E–F) CSCs, MEK1 inhibition CSCs and SIRT1 overexpression while MEK1 inhibition CSCs were prepared for 14 days, then subcutaneous injected in NOD-SCID mice (CSCs control group 4 mice, other two groups 8 mice each). Tumor sizes were measured with calipers in three dimensions every other day. Tumor volumes were calculated using the formula: tumor volume (cm3) = 0.52 × (W) 2 × (L), where L is length and W is width. We counted and weight the tumors, 30 days later.